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In situPCR on Plant Material with Sub-cellular Resolution

Authors
Journal
Annals of Botany
0305-7364
Publisher
Oxford University Press
Publication Date
Volume
80
Issue
5
Identifiers
DOI: 10.1006/anbo.1997.0502
Keywords
  • In Situpcr
  • Reversed Transcribedin Situpcr
  • Rbcl
  • C4-Plants

Abstract

Abstract In situPCR and reverse transcribed in situPCR have been tested on leaves of sugar cane fixed in FAA, 4% PFA or 2% PFA+2.5% GA. In situPCR amplification of the gene coding for ribulose-1,5-bisphosphate carboxylase/oxygenase ( rbcL) was successfully performed following all three fixation protocols. As expected the PCR product was restricted to the plastids of all cells. Reverse transcribed in situPCR was performed on rbcL mRNA and in this case the PCR product was restricted to the plastids of the bundle sheath cells. This is the first report of in situPCR on plant material and only the second report of in situPCR with sub-cellular resolution. In situPCR on rbcL may prove to be a valuable positive control for future in situPCR studies on plant material.

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